Molecular cloning, in vitro expression and bioactivity of rabbit transforming growth factor-beta receptor type II (rTGF-βRII)

Vet Immunol Immunopathol. 2011 Mar 15;140(1-2):55-62. doi: 10.1016/j.vetimm.2010.11.014. Epub 2010 Nov 30.

Abstract

Transforming growth factor-beta receptor II (TGF-βRII) is an attractive target for anti-scarring therapy in wound healing because it attenuates excessive TGF-β which has pleiotropic effects on the immune system. In the present study, the cDNA of rabbit TGF-βRII (rTGF-βRII) was amplified from rabbit peripheral blood by RT-PCR. The open reading frame of rTGF-βRII encodes a protein consisting of 567 amino acids, which contains a predicted transmembrane domain and a Serine/Threonine protein kinase domain similar to other identified mammalian TGF-βRIIs. The amino acid sequences of the biologically active, soluble rTGF-βRII and mouse, rat, human and chicken counterparts are 81%, 81%, 89% and 61%, respectively, identical. Recombinant soluble rTGF-βRII (rsTGF-βRII) fused with His tag was efficiently expressed in Escherichia coli BL21 (DE3) expression host strain. This fusion protein's molecular weight of ∼ 19 kDa was identified by SDS-PAGE and Western blotting. In vitro, purified rsTGF-βRII was able to inhibit the proliferation of keloid rabbit fibroblasts and decrease the level of collagen. These findings indicate that rTGF-βRII plays an important role in inhibiting the proliferation of keloid rabbit fibroblasts and provides the basis for investigations on the role of TGF-βRII in this important domestic species.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Western / veterinary
  • Cloning, Molecular*
  • Fibroblasts / metabolism
  • In Vitro Techniques
  • Keloid / metabolism
  • Male
  • Molecular Sequence Data
  • Protein Serine-Threonine Kinases* / biosynthesis
  • Protein Serine-Threonine Kinases* / genetics
  • Protein Serine-Threonine Kinases* / metabolism
  • Rabbits / genetics*
  • Rabbits / immunology
  • Receptor, Transforming Growth Factor-beta Type II
  • Receptors, Transforming Growth Factor beta* / biosynthesis
  • Receptors, Transforming Growth Factor beta* / genetics
  • Receptors, Transforming Growth Factor beta* / metabolism
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction / veterinary
  • Sequence Analysis, DNA / veterinary
  • Wound Healing*

Substances

  • Receptors, Transforming Growth Factor beta
  • Recombinant Fusion Proteins
  • Protein Serine-Threonine Kinases
  • Receptor, Transforming Growth Factor-beta Type II

Associated data

  • GENBANK/GU987103